Protein ID (NP or XP #) or Wolbachia#:
Tb927.10.6690
Organism (including strain): Trypanosoma brucei
Etiologic Risk Group (see link below):
Risk Group 2
Background/Disease Information: Trypanosoma brucei are microscopic parasites that causes African Trypanosomiasis, also known as "sleeping sickness." The parasite makes its way into the blood-brain barrier and infects the central nervous system causing fever, headache, muscle and joint aches, enlarged lymph nodes and sores around the wound. As time progresses, it may cause mental deterioration and other neurologic problems. Death ensues usually within a few months. It is transmitted by the tsetse fly (Glossina species), which is found only in rural Africa. Although the infection is not found in the United States, it is a serious public health problem in some regions of sub-Saharan Africa. Sleeping sickness is curable with medication, but if left untreated, the infection will eventually lead to coma and death.
Essentiality of this protein:
Tb927.10.6690 has essentiality data Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in bloodstream forms (3 days); Source study: alsford Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in bloodstream forms (6 days); Source study: alsford Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in procyclic forms; Source study: alsford Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in differentiation of procyclic to bloodstream forms; Source study: alsford
Complex of proteins:
None
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
Druggability index (range: 0 to 1): 0.3
No chemical compounds associated to this gene
No orthologous druggable targets
No additional associated druggable targets
List cost and quantity of substrate reagents, supplier, and catalog #:
Substrate
Cost
Quantity
Supplier
Catalog #
4-nitrophenyl phosphate
209.50
50
Sigma-Aldrich Corporation
333338-18-4
Raytide peptide
Contact
Contact
Manufactured by EMD Biosciences, Inc
PK02
Structure Available (PDB or Homology model): 3M4U
Current Inhibitors:
H3VO4
Expression Information (has it been expressed in bacterial cells):
Escherichiu coli
Purification Method:
Purification of plasma membranes described by Seyfang (Scyfdng and Duszenko, 1993) and fractions were stored at -80°C. Briefly. the ghost fraction (F2) was stripped of peripheral and cytoskeletal proteins by EDTA/alkali treatment (F3), and octylthioglucoside was used to solubilize plasma-membrane proteins (F4). The protein concentration was determined by the method of Bradford (1976) using BSA as the standard and 0.1% Triton X-100 to solubilize membrane proteins.
Length of your protein in Amino Acids:
Sequence Length: 306 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website
34132.3
Molar Extinction coefficient of your protein at 280 nm wavelength:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 29005 Abs 0.1% (=1 g/l) 0.850, assuming all pairs of Cys residues form cystines
Ext. coefficient 28880 Abs 0.1% (=1 g/l) 0.846, assuming all Cys residues are reduced
Trypanosoma brucei, protein tyrosine phosphatase
NCBI Gene # or RefSeq#:
295789515
http://www.ncbi.nlm.nih.gov/protein/3M4U_A
Protein ID (NP or XP #) or Wolbachia#:
Tb927.10.6690
Organism (including strain):
Trypanosoma brucei
Etiologic Risk Group (see link below):
Risk Group 2
Background/Disease Information:
Trypanosoma brucei are microscopic parasites that causes African Trypanosomiasis, also known as "sleeping sickness." The parasite makes its way into the blood-brain barrier and infects the central nervous system causing fever, headache, muscle and joint aches, enlarged lymph nodes and sores around the wound. As time progresses, it may cause mental deterioration and other neurologic problems. Death ensues usually within a few months. It is transmitted by the tsetse fly (Glossina species), which is found only in rural Africa. Although the infection is not found in the United States, it is a serious public health problem in some regions of sub-Saharan Africa. Sleeping sickness is curable with medication, but if left untreated, the infection will eventually lead to coma and death.
Link to TDR Targets page:
http://www.tdrtargets.org/targets/view?gene_id=10415
Link to Gene Database page:
http://www.genedb.org/gene/Tb927.10.6690
Essentiality of this protein:
Tb927.10.6690 has essentiality data
Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in bloodstream forms (3 days); Source study: alsford
Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in bloodstream forms (6 days); Source study: alsford
Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in procyclic forms; Source study: alsford
Gene/Ortholog: Tb927.10.6690 (OG4_23831); Phenotype: no significant loss or gain of fitness in differentiation of procyclic to bloodstream forms; Source study: alsford
Complex of proteins:
None
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
Druggability index (range: 0 to 1): 0.3
No chemical compounds associated to this gene
No orthologous druggable targets
No additional associated druggable targets
EC#:
3.1.3.48
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/php/result_flat.php4?ecno=3.1.3.48
Enzyme Assay Information Link:
http://www.brenda-enzymes.info/literature/lit.php4?e=3.1.3.48&r=95003
http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1995.871_a.x/pdf
List cost and quantity of substrate reagents, supplier, and catalog #:
Structure Available (PDB or Homology model):
3M4U
Current Inhibitors:
H3VO4
Expression Information (has it been expressed in bacterial cells):
Escherichiu coli
Purification Method:
Purification of plasma membranes described by Seyfang (Scyfdng and Duszenko, 1993) and fractions were stored at -80°C. Briefly. the ghost fraction (F2) was stripped of peripheral and cytoskeletal proteins by EDTA/alkali treatment (F3), and octylthioglucoside was used to solubilize plasma-membrane proteins (F4). The protein concentration was determined by the method of Bradford (1976) using BSA as the standard and 0.1% Triton X-100 to solubilize membrane proteins.
Image of protein:
Amino Acid Sequence:
MSTAKSFPMAQLSTRAQYSRMQREFVQLQRQENPRNINFTTSLKNRHKNRYLDILANEET
IYPPVLKAVGAQPGRYPYINGNLIDLDLPHTFVACQAPVPQGVPDFLETLSEKKVDLVVM
LTKLREGGVLKAERYWPEEEEDSLSFPESGHDAIKVTRDAEASYEVDAELDIVRRPLVIH
VPGKPMHRVLQVQYVGWPDHGVPESAASFDELLSVIKNCVTTSPILVHCSAGIGRTGTLI
GAYAALLHIERGILTDSTVYSIVAAMKQKRFGMVQRLEQYAVIYMTVLGRLGVDISGLVS
TLNLKA
Length of your protein in Amino Acids:
Sequence Length: 306 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website
34132.3
Molar Extinction coefficient of your protein at 280 nm wavelength:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 29005 Abs 0.1% (=1 g/l) 0.850, assuming all pairs of Cys residues form cystines
Ext. coefficient 28880 Abs 0.1% (=1 g/l) 0.846, assuming all Cys residues are reduced
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
CDS Gene Sequence (paste as text only):
GC% Content for gene:
CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
GC% Content for gene (codon optimized):
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
Primer design results for 'tail' primers (this is just 2 sequences):
Resources:
See ProtocolTargetDiscoveryVDS.docx for more
Etiologic Risk Group Categories (for pathogens): http://www.utexas.edu/research/rsc/ibc/agent_class.html#_Toc7238334
SIGMA-ALDRICH RESOURCES
Enzyme Explorer
http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer.html
Enzyme Classification Index (EC number)
http://www.sigmaaldrich.com/life-science/biochemicals/biochemical-products.html?TablePage=14573088
WolframAlpha http://www.wolframalpha.com/
DrugBank http://www.drugbank.ca/
Databases of genes/organisms:
http://www.niaid.nih.gov/Pages/default.aspx
http://eupathdb.org/eupathdb/
https://patricbrc.vbi.vt.edu/portal/portal/patric/Home
http://www.nmpdr.org/FIG/wiki/view.cgi/Main/EssentialGenes
http://tubic.tju.edu.cn/deg/
http://csgid.org/csgid/cake/pages/community_request_gateway
http://tdrtargets.org/
http://gsc.jcvi.org/status.shtml
Scientific Nomenclature page from Center for Disease Control (gene, protein names and abbreviations)
http://wwwnc.cdc.gov/eid/pages/scientific-nomenclature.htm
Gene Information:
NCBI GENE Page: http://www.ncbi.nlm.nih.gov/gene
BLAST Page: http://blast.ncbi.nlm.nih.gov/
Protein Information:
NCBI Protein Page: http://www.ncbi.nlm.nih.gov/protein
Protein Expression Website
Protein Expression Paper: SGC_ProteinProductionPurificationNatMethods2008.pdf
Primer Overlap PCR Articles
HooverLubkowski_PCRoverlapcloninggnf042.pdf
StemmerPCRoverlapGene1995.pdf
Is my target good for Virtual Screening programs?
Reynolds_THermodynamicsLigandBinding_MedChemLett2011.pdf